sparQ DNA Library Prep Kit

sparQ DNA Library Prep Kit

Streamlined, versatile single-tube solution for high quality library prep

Features & Benefits

Fast, easy single-tube solution completes library prep in 2.5 hours
Suitable for a wide range of input amounts from as low as 250 pg
Optimized chemistry ensuring superior library prep sensitivity and efficiency
Higher library yields compared to other library prep kits
High efficiency enables PCR-free workflow from 100 ng input

sparQ DNA Library Prep Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.

Description

sparQ DNA Library Prep Kit is optimized for the rapid construction of DNA libraries from fragmented double-stranded DNA for sequencing on Illumina® NGS platforms. The simplified protocol speeds up library prep to 2.5 hours with minimal hands-on time and accommodates DNA input amounts from 250 pg to 1 μg. DNA polishing reactions are streamlined into a single step to convert fragmented DNA into 5'-phosphorylated and 3'-dA-tailed DNA fragments. This is followed by high efficiency adapter ligation in the same tube. PCR-free workflows are enabled from 100 ng of starting material. If library amplification is required, the HiFi PCR Master Mix and Primer Mix ensure even amplification with minimal bias.

Streamlined workflow

sparQ DNA Library Prep workflow

Generates sequencing ready library in 2.5 hours with minimal hands-on time

Simplified single-tube solution minimizes sample loss

Higher library yields

Higher library yields

Higher library yields

Higher library yields

sparQ DNA Library Prep Kit produces high quality libraries from a broad range of DNA inputs with significantly higher yields. Libraries were prepared with Covaris-sheared human genomic DNA (250 bp average size) using kit manufacturers’ instructions. Amplified libraries (6 PCR cycles for 100 ng input DNA and 13 PCR cycles for 1 ng input DNA) were quantified with Qubit fluorometric method.

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Unique enzymes engineered for optimal sensitivity and efficiency

Over 50% increase in library yields than other library prep kits

Compatible with a broad range of DNA inputs from 250 pg to 1 μg

Uniform coverage across challenging regions

Coverage across a wide range of GC-content

Coverage across a wide range of GC-content

Coverage across a wide range of GC-content

PCR-free (dark blue) and amplified libraries (orange) were prepared from 100 ng of microbial genomic DNA of different species using sparQ DNA Library Prep Kit. Libraries were sequenced on Illumina MiSeq and 2 million sequencing reads from each library was analyzed. Coverage uniformity was examined by plotting normalized coverages of the library against GC-content of the targeted genome. GC-content distribution of targeted genomes is indicated by gray line. Both PCR-free and amplified libraries resulted in uniform coverage across a wide range of GC-content..

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Even coverage across a broad GC-content

Coverage of amplified libraries closely resembles coverage of PCR-free workflow

High quality sequencing metrics

Higher quality sequencing metrics

Higher quality sequencing metrics

Higher quality sequencing metrics

sparQ DNA Library Prep Kit generates high quality DNA libraries with minimal duplication rates. Libraries were prepared with 1 ng and 100 ng of microbial genomic DNA and subsequently sequenced on Illumina MiSeq. Each library was down-sampled to 2 million reads (150 bp paired-end reads) and aligned to a reference genome with only unique alignments reported.

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High mapping percentage and low duplication artifacts

Industry leading sequencing results ensures better return on investments

NSG Automation

Quantabio sparQ Frag & Library Prep Kit can be fully automated

Currently sparQ Frag & Library Prep Kit is supported by the following vendors:

Looking for more information or for different automation vendor?
Contact Us

Vendor Automation Platform(s)

PerkinElmer Applied Genomics SCICLONE G3 NGSx

Beckman Coulter in development

Tecan in development

sparQ UDI Adapter Resources

Setting up sparQ Library Prep kits in Illumina Experiment Manager

Technical Note - Setting up sparQ Library Prep Kit in Illumina Experiment Manager

Illumina Experiment Manager version 1.8 files (zip-download)

Illumina Experiment Manager version 1.9 files (zip-download)

sparQ UDI Adapter sequence overview

Vendor	Automation Platform(s)
PerkinElmer Applied Genomics	SCICLONE G3 NGSx
Beckman Coulter	in development
Tecan	in development

Performance Data

sparQ DNA Library Prep Workflow

sparQ DNA Library Prep Workflow

sparQ DNA Library Prep Workflow

The streamlined workflow can be completed in under 3 hours with minimal hands-on time. A single tube is used for DNA polishing, ligation, and cleanup. A second tube is used for workflows requiring PCR amplification and a final tube receives the sequencing-ready library.

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Coverage Across Wide Range of GC-Content

Coverage Across Wide Range of GC-Content

Uniform coverage across wide range of GC-content

Library amplification with sparQ HiFi PCR Master Mix contained in the sparQ DNA Library Prep Kit resulted in uniform coverage across the wide range of GC-content. Libraries were prepared by using sparQ DNA Library Prep Kit with 100 ng input DNA. Coverage depth against GC-content of libraries amplified by sparQ HiFi PCR Master Mix (orange) were compared to corresponding libraries without amplification (dark blue: PCR-free library). GC content distribution of targeted genomes is indicated by gray line.

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Improve Sequencing Results and Economics

Improve Sequencing Results and Economics

Improve Sequencing Results and Economics

sparQ DNA Library Prep Kit generates high quality DNA libraries with minimal duplication rates. Libraries were prepared with 1 ng and 100 ng of microbial genomic DNA and subsequently sequenced on Illumina MiSeq. Each library was down-sampled to 2 million reads (150 bp paired-end reads) and aligned to a reference genome with only unique alignments reported.

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Coverage Across Challenging Regions

Coverage Across Challenging Regions

Coverage Across Challenging Regions

Amplified libraries were prepared from 100 ng of microbial genomic DNA and subsequently sequenced on Illumina MiSeq. 2 million reads from each tested library were down-sampled and analyzed. Coverage uniformity for different library preparation kits were compared by plotting normalized coverage for both extreme AT-rich regions (8%-20% GC-content) and GC-rich regions (75%-88% GC-content).

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sparQ DNA Library Prep Kit Yield Analysis

sparQ DNA Library Prep Kit Yield Analysis

sparQ DNA Library Prep Kit Yield Analysis

sparQ DNA Library Prep Kit produces high quality libraries from a broad range of DNA inputs with significantly higher yields. Libraries were prepared with Covaris-sheared DNA (250 bp average size) using kit manufacturers’ instructions. Amplified libraries (6 amplification cycles for 100 ng input DNA and 13 amplification cycles for 1 ng input DNA) were quantified with Qubit fluorometric quantitation method.

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sparQ UDI Adapter Resource
For help setting up UDI’s in Illumina Experiment Manager Download

sparQ NGS Library Prep Solutions

Download Poster

Streamlined single-tube solutions for high quality DNA library preparation

Details

Contents

Volume

Component Description

Cap Color

95191-024

95191-096

DNA Polishing Enzyme Mix

Blue

1 x 0.24 ml

1 x 0.96 ml

DNA Polishing Buffer

Blue

1 x 0.12 ml

1 x 0.48 ml

DNA Ligase

Orange

1 x 0.24 ml

1 x 0.96 ml

DNA Rapid Ligation Buffer

Orange

1 x 0.48 ml

2 x 0.96 ml

HiFi PCR Master Mix (2X)

White

1 x 0.60 ml

2 x 1.25 ml

Primer Mix

White

1 x 0.036 ml

1 x 0.144 ml

Storage & Handling

Store kit components in a constant temperature freezer at -25°C to -15°C upon receipt. For lot specific expiry date, refer to package label, Certificate of Analysis or Product Specification Form.

Related Resources

Product Flyers

sparQ DNA Library Prep Flyer

Product Manuals

sparQ DNA Library Prep Kit

Technical Notes

Streamlined single-tube solutions for high quality DNA library preparation Setting up sparQ Library Prep Kit in Illumina Experiment Manager

Publications

Detailed temporal dissection of an enhancer cluster reveals two distinct roles for individual elements
Henry Thomas, bioRxiv - 2020
Abstract
Many genes are regulated by multiple enhancers that often simultaneously activate their target gene. Yet, how individual enhancers collaborate to activate transcription is not well understood. Here, we dissect the functions and interdependencies of five enhancer elements that form a previously identified enhancer cluster and activate the Fgf5 locus during exit from naïve murine pluripotency. Four elements are located downstream of the Fgf5 gene and form a super-enhancer. Each of these elements contributes to Fgf5 induction at a distinct time point of differentiation. The fifth element is located in the first intron of the Fgf5 gene and contributes to Fgf5 expression at every time point by amplifying overall Fgf5 expression levels. This amplifier element strongly accumulates paused RNA Polymerase II but does not give rise to a mature Fgf5 mRNA. By transplanting the amplifier to a different genomic position, we demonstrate that it enriches for high levels of paused RNA Polymerase II autonomously. Based on our data, we propose a model for a mechanism by which RNA Polymerase II accumulation at a novel type of enhancer element, the amplifier, contributes to enhancer collaboration.
Genomic Insights and Ecological Adaptations of Deep-Subsurface and Near Subsurface Thermococcus Isolates
Lilja Caitlin Strang, Western CEDAR - 2020
Abstract
Members of the Archaeal genus Thermococcus are sulfur-dependent hyperthermophiles found in hydrothermal vents throughout the world. Previous analysis of a Thermococcus culture collection containing isolates from the Juan de Fuca Ridge, Gorda Ridge, and South East Pacific Rise using amplified fragment length polymorphism analysis and multilocus sequence typing revealed a distinct clade of Thermococcus isolated from the 1996 megaplume event at Gorda Ridge, indicating that they originated from a deep-subsurface habitat. The aim of this study was to elucidate the functional adaptations that allow for the survival of the Gorda Ridge clade in a deepsubsurface habitat as compared to representative Thermococcus isolates from shallow subsurface environments. This was accomplished through a pangenomic analysis of representative isolates in this clade and others from this culture collection. The Gorda Ridge megaplume group was enriched for genes relating to DNA repair and stabilization including a predicted endonuclease distantly related to Archaeal Holliday junction resolvase, DNA mismatch repair ATPase mutS, CRISPR/Cas elements, and dnaK (hsp70). The group was also enriched for ABC-type branched-chain amino acid (BCAA) transport system, enzymes for the Shikimate pathway for aromatic amino acid synthesis, as well as TupA for tungstate transport. These findings suggest that Thermococcus inhabiting deep-subsurface fluid reservoir require the added ability to prevent and repair damage to their DNA, presumably due to the energy demands of DNA replication. The enrichment in BCAA and tungstate transporters may indicate the use of an amino acid catabolism pathway followed by fermentation catalyzed by the tungstopterin containing enzymes aldehyde ferredoxin oxidoreductase and alcohol dehydrogenase, suggesting a preference for peptides over carbohydrates as an energy source in the deep-subsurface.
Draft Genome Sequence of Phyllobacterium endophythicum mTS5, Isolated from Lupinus micranthus in Tunisia
Zoé Waller, Microbiology Resource Announcements - 2020
Abstract
We report here the draft genome sequence of Phyllobacterium endophyticum mTS5, isolated from a Lupinus micranthus root nodule. The genome consists of 5,454,168 bp, with a GC content of 57%, and contains 5,676 protein-coding sequences.
Click here to see all Publications

Safety Data Sheets (SDS)

sparQ DNA Library Prep SDS HiFi PCR Master Mix sparQ DNA Library Prep SDS DNA Ligation Buffer sparQ DNA Library Prep SDS DNA Polishing Buffer sparQ DNA Library Prep SDS DNA Polishing Enzyme sparQ DNA Library Prep SDS Primer Mix sparQ DNA Library Prep SDS T4 DNA Ligase

CofA (PSF)

PSF-95191-024-Lot#022276 PSF-95191-096-Lot#022277 PSF-95191-024-Lot#024784 PSF-95191-096-Lot#024785 PSF-95191-024-Lot#026594 PSF-95191-024-Lot#027775 PSF-95191-096-Lot#027826 PSF-95191-024 -Lot#029537 PSF-95191-096 -LOT#66149997 PSF-95191-024-LOT#66153845 PSF-95191-024-LOT#66168084 PSF-95191-024-Lot#66168084 PSF-95191-024-Lot#66169235 PSF-95191-096-Lot#66169243

Testimonials

sparQ DNA Library Prep Kit

"Very easy to use."

Scientist

University Sherbrooke

Ordering Information

Product

Kit Size

Order Info

sparQ DNA Library Prep Kit
Request Sample

24 rxns

Order (95191-024)

96 rxns

Order (95191-096)

sparQ UDI Adapters (1-96)

1 - 96

Order (95211-096)

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5 mL

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450 mL

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		Volume
Component Description	Cap Color	95191-024	95191-096
DNA Polishing Enzyme Mix	Blue	1 x 0.24 ml	1 x 0.96 ml
DNA Polishing Buffer	Blue	1 x 0.12 ml	1 x 0.48 ml
DNA Ligase	Orange	1 x 0.24 ml	1 x 0.96 ml
DNA Rapid Ligation Buffer	Orange	1 x 0.48 ml	2 x 0.96 ml
HiFi PCR Master Mix (2X)	White	1 x 0.60 ml	2 x 1.25 ml
Primer Mix	White	1 x 0.036 ml	1 x 0.144 ml